Abstract
Proteins are dynamic molecules that can undergo rapid conformational rearrangements in response to stimuli. These structural changes are often critical to protein function, and thus elucidating time-dependent conformational landscapes has been a long-standing goal of structural biology. To harness the power of single particle cryo-EM methods to enable ‘time-resolved’ structure determination, we have developed a light-coupled cryo-plunger that pairs flash-photolysis of caged ligands with rapid sample vitrification. The ‘flash-plunger’ consists of a high-power ultraviolet LED coupled with focusing optics and a motorized linear actuator, enabling the user to immobilize protein targets in vitreous ice within a programmable time window – as short as tens of milliseconds – after stimulus delivery. The flash-plunger is a simple, inexpensive and flexible tool to explore short-lived conformational states previously unobtainable by conventional sample preparation methods.
Original language | English (US) |
---|---|
Article number | 107624 |
Journal | Journal of Structural Biology |
Volume | 212 |
Issue number | 3 |
DOIs | |
State | Published - Dec 1 2020 |
Keywords
- Cryo-EM
- Cryo-electron microscopy
- Manual cryo-plunger
- Single particle
- Time-resolved
ASJC Scopus subject areas
- Structural Biology